Genome-wide association studies have identified autophagy-related susceptibility genes for inflammatory bowel disease (); however, whether autophagy regulators can be utilized as therapeutic targets remains unclear. To identify novel microtubule-associated protein 1 light chain 3 (LC3)-interacting proteins in intestinal epithelial cells (IECs), we isolated primary IECs from green fluorescent protein (GFP)-LC3 mice. We performed immunoprecipitation with a GFP antibody and then analyzed co-immunoprecipitates by mass spectrometry. HADHA was identified as an LC3-interacting protein from primary IECs. The HADHA gene encodes the alpha subunit of the mitochondrial trifunctional protein. Given that HADHA catalyzes the last three steps of mitochondrial beta-oxidation of long-chain fatty acids, we investigated whether long-chain fatty acids induce autophagy in IECs. We found that induced autophagy in DLD-1, HT29, and HCT116\xa0cells. HADHA was expressed in not only the mitochondria but also the cytosol. LC3 puncta co-localized with HADHA, which were enhanced by stimulation. However, LC3 puncta did not co-localize with Tom20, suggesting that HADHA was induced to associate with LC3 puncta at sites other than the mitochondria. Thus, HADHA may have extra-mitochondrial functions. Furthermore, we found that induced cell death in IECs, which was accelerated by bafilomycin A and chloroquine. These findings suggested that -induced autophagy supports the survival of IECs. Taken together, these results suggested that HADHA is involved in long-chain fatty -induced autophagy in IECs, thus providing new insights into the pathology of and revealing novel therapeutic targets of .Copyright © 2017 Elsevier Inc. All rights reserved.
Keyword: IBD
Structural studies of the major glycolipids produced by two Lactobacillus johnsonii (LJ) strains, LJ 151 isolated from intestinal tract of healthy mice and LJ 142 isolated from mice with experimentally induced inflammatory bowel disease (), were performed. Two major glycolipids, GL1 and GL2, were present in lipid extracts from L.\xa0johnsonii 142 and 151 strains. Glycolipid GL1 has been identified as β-D-Glcp-(1→6)-α-D-Galp-(1→2)-α-D-Glcp-diglyceride and GL2 as α-D-Galp-(1→2)-α-D-Glcp-diglyceride. The main fatty residues identified by gas-liquid chromatography-mass spectrometry were , stearic and lactobacillic acids. Besides structural elucidation of the major glycolipids, the aim of this study was to determine the immunochemical properties of these glycolipids and to compare their immunoreactivity to that of polysaccharides obtained from the same strains. Sera from rabbits immunized with bacterial cells possessed much higher serological reactivity with polysaccharides than with glycolipids. Inversely, reactivity of the glycolipids with human sera from patients with was much higher than that determined for the polysaccharides, while reactivity of glycolipids with human sera from healthy individuals was much lower than one measured for the polysaccharides. Results indicate that glycoconjugates from Lactobacillus cell wall act as antigens and may represent new diagnostic biomarkers.© 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.
Keyword: IBD
A new class of outer membrane lipid (OML) was isolated from the oral spirochete Treponema denticola strain ATCC 33521 using a phenol/chloroform/light petroleum procedure normally applied for lipopolysaccharide extraction. In addition to chemical analysis, Fourier transform infrared (FTIR) spectroscopy was applied to compare the biophysical properties of OML with lipopolysaccharides (LPS) and lipoteichoic acids (LTA). Isolated OML fractions represent 1.4% of the total dry cell weight, are about 4 kDa in size, and contain 6% amino sugars, 8% neutral sugars, 14% phosphate, 35% carbazol-positive compounds, and 11% fatty acids (containing iso- and anteiso-fatty acyl chains). Rare for outer membrane lipids, OML contains no significant amount of 3-deoxy-D-manno-octulosonic acids, heptoses, and beta-hydroxy fatty acids. The fatty acyl chain composition, being similar to that of the cytoplasmic membrane, is quite heterogeneous with anteiso-pentadecanoic (12%), (51%), and iso- (19%) as the predominant fatty acids present. Findings of a glycerol-hexose unit and two glycerol-hexadecanoic fragments indicate a glycolipid membrane anchor typically found in LTA. There was also no evidence for the presence of a sphingosine-based lipid structure. The results of FTIR measurements strongly suggest that the reconstituted lipid forms normal bilayer structures (vesicles) expressing a high membrane state of order with a distinct phase transition as typical for isolated LPS. However, in contrast to LPS, OML of T. denticola has a lower Tm near 22 degreesC and a lower cooperativity of the phase transition. The results suggest a different kind of permeation barrier that is built up by this particular OML of T. denticola, which is quite different from LPS normally essential for Gram-negative bacteria.
Keyword: IBD
Fatty amides (FAAs), conjugates of fatty acids with ethanolamine, mono-amine neurotransmitters or amino acids are a class of molecules that display diverse functional roles in different cells and tissues. Recently we reported that one of the serotonin-fatty conjugates, docosahexaenoyl serotonin (DHA-5-HT), previously found in gut tissue of mouse and pig, attenuates the IL-23-IL-17 signaling axis in LPS-stimulated mice macrophages. However, its presence and effects in humans remained to be elucidated. Here, we report for the first time its identification in human intestinal (colon) tissue, along with a series of related N-acyl serotonins. Furthermore, we tested these fatty conjugates for their ability to inhibit the release of IL-17 and CCL-20 by stimulated human peripheral blood mononuclear cells (PBMCs). Serotonin conjugates with (PA-5-HT), stearic (SA-5-HT) and oleic (OA-5-HT) were detected in higher levels than arachidonoyl serotonin (AA-5-HT) and DHA-5-HT, while eicosapentaenoyl serotonin (EPA-5-HT) could not be quantified. Among these, DHA-5-HT was the most potent in inhibiting IL-17 and CCL-20, typical Th17 pro-inflammatory mediators, by Concanavalin A (ConA)-stimulated human PBMCs. These results underline the idea that DHA-5-HT is a gut-specific endogenously produced mediator with the capacity to modulate the IL-17/Th17 signaling response. Our findings may be of relevance in relation to intestinal inflammatory diseases like Crohn's disease and Ulcerative colitis.Copyright © 2017. Published by Elsevier B.V.
Keyword: IBD
Wavelength of light is a crucial factor which renders microalgae as the potential biodiesel. In this study, Tetraselmis sp. and Nannochloropsis sp. as famous targets were selected. The effect of different light wavelengths on growth rate and lipid production was studied. Microalgae were cultivated for 14 days as under blue, red, red-blue LED and white fluorescent light. The growth rate of microalgae was analyzed by spectrophotometer and cell counting while oil production under improved Nile red method. Optical density result showed the microalgae exhibited better growth curve under blue wavelength. Besides, Tetraselmis sp. and Nannochloropsis sp. under blue wavelength showed the higher growth rate (1.47 and 1.64 day(-1)) and oil production (102.954 and 702.366 a.u.). Gas chromatography analysis also showed that and stearic which were compulsory components for biodiesel contribute around 49-51% of total FAME from Nannochloropsis sp. and 81-83% of total FAME from Tetraselmis sp.Copyright © 2014 Elsevier Ltd. All rights reserved.
Keyword: IBD
Fatty acids and their derivatives play an important role in inflammation. Diet and genetics influence fatty profiles. Abnormalities of fatty profiles have been observed in inflammatory bowel diseases (), a group of complex diseases defined by chronic gastrointestinal inflammation. associated fatty profile abnormalities were observed independently of nutritional status or disease activity, suggesting a common genetic background. However, no study so far has attempted to look for overlap between loci and fatty associated loci or investigate the genetics of fatty profiles in . To this end, we conducted a comprehensive genetic study of fatty profiles in using iCHIP, a custom microarray platform designed for deep sequencing of immune-mediated disease associated loci. This study identifies 10 loci associated with fatty profiles in . The most significant associations were a locus near CBS (p\u202f=\u202f7.62\u202f×\u202f10) and a locus in LRRK2 (p\u202f=\u202f1.4\u202f×\u202f10). Of note, this study replicates the FADS gene cluster locus, previously associated with both fatty profiles and pathogenesis. Furthermore, we identify 18 carbon chain trans-fatty acids (p\u202f=\u202f1.12\u202f×\u202f10), total trans-fatty acids (p\u202f=\u202f4.49\u202f×\u202f10), (p\u202f=\u202f5.85\u202f×\u202f10) and arachidonic (p\u202f=\u202f8.58\u202f×\u202f10) as significantly associated with pathogenesis.Copyright © 2018 Elsevier Ltd. All rights reserved.
Keyword: IBD
N-Palmitoylethanolamine or palmitoylethanolamide (PEA) is an anti-inflammatory compound that was recently shown to exert peroxisome proliferator-activated receptor-α-dependent beneficial effects on colon inflammation. The actions of PEA are terminated following hydrolysis by 2 enzymes: fatty amide hydrolase (FAAH), and the less-studied N-acylethanolamine-hydrolyzing amidase (NAAA). This study aims to investigate the effects of inhibiting the enzymes responsible for PEA hydrolysis in colon inflammation in order to propose a potential therapeutic target for inflammatory bowel diseases (IBDs). Two murine models of were used to assess the effects of NAAA inhibition, FAAH inhibition, and PEA on macroscopic signs of colon inflammation, macrophage/neutrophil infiltration, and the expression of proinflammatory mediators in the colon, as well as on the colitis-related systemic inflammation. NAAA inhibition increases PEA levels in the colon and reduces colon inflammation and systemic inflammation, similarly to PEA. FAAH inhibition, however, does not increase PEA levels in the colon and does not affect the macroscopic signs of colon inflammation or immune cell infiltration. This is the first report of an anti-inflammatory effect of a systemically administered NAAA inhibitor. Because NAAA is the enzyme responsible for the control of PEA levels in the colon, we put forth this enzyme as a potential therapeutic target in chronic inflammation in general and in particular.© FASEB.
Keyword: IBD